MicroCal iTC200 and Auto-iTC200 instruments (GE Healthcare) were used to perform ITC measurements as described previously (Julien et al., 2013b (link)). For the PGT151:BG505 SOSIP.664 titration experiment, the Env trimer was deposited in the cell at concentrations between 4.4 and 4.7 μM, and the Fab was in the pipette at concentrations between 79 and 102 μM. In the glycan: PGT151 titration experiments, PGT151 Fab was added to the cell at 100 μM with glycans in the pipette at 4-5 mM (for synthesis of glycans, see Supplemental Experimental Procedures ). Data analysis was carried out with Origin 7.0 software using a single-site binding model.
Auto itc200
Manufactured by GE Healthcare
Sourced in United States
The Auto-iTC200 is a fully automated, high-throughput isothermal titration calorimetry (ITC) system designed for the accurate and efficient measurement of biomolecular interactions. The instrument automatically performs sample preparation, titration, and data analysis, providing reliable and reproducible results.
Automatically generated - may contain errors
Lab products found in correlation
18 protocols using auto itc200
1
Thermodynamic Analysis of Env Trimer Binding
2
DAZAP1 Binding Kinetics with hnRNPs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ITC experiment was performed on auto ITC 200 (GE Health care). The purified recombinant proteins were resuspended in 20 mM Na-Phosphate buffer pH 7.0 with 150 mM NaCl for ITC analysis. The full length or truncated DAZAP1 (100–150 μM in the injection syringe) was titrated into other hnRNPs (10–15 μM of hnRNPs in the cell). Injections of 2 μl DAZAP1 protein for 20 titration points were performed and the data was fitted using Origin for ITC software. All experiments were carried out in duplicates and the average experiments were reported.
3
Isothermal Titration Calorimetry of Beclin1-Bcl2 Interaction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isothermal Titration Calorimetry was performed using Auto-iTC200 (GE, USA). Samples were dialysed into 50 mM Tris, pH 8.0, and 150 mM NaCl before experiment. To study the interaction between Beclin1 and Bcl2 in the absence of MA, we used a typical titration consisting of injecting 2 μl aliquots of 200 μM Beclin1 solution into 200 μl aliquots of 20 μM Bcl2 solution after every 1 min to ensure the titration peak would return to the baseline prior to the next injection. Aliquots of the same Beclin1 solutions were injected into only the reaction buffer (50 mM Tris containing 150 mM NaCl, pH 8.0) in separate ITC runs to measure the heats of dilution of the ligands. To study the interaction between Beclin1 and Bcl2 in the presence of MA, 200 μM Beclin1 solution was titrated into 200 μl aliquots of 20 μM Bcl2 solution containing 10 μM MA with the settings as above. Control experiments were performed by titrating Beclin1 solution into the same buffer to obtain the heats of dilution. Typically, titrations consisted of 20 injections of 2 μl, with 180-s equilibration between injections. Each reaction was repeated 3 times. The data were analysed using Origin 7.0.
4
Glycan-Protein Binding Affinity by ITC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An Auto-iTC200 instrument (GE Healthcare) was used to perform isothermal titration calorimetry (ITC). O6 recombinant protein was purified, and lyophilized glycans were resuspended, in a buffer containing 25 mM HEPES pH 7.5 and 250 mM sodium chloride. Glycans were obtained from the CFG or purchased from Glycotech or Sigma and contained an Sp-biotin linker (CH2CH2NH-biotin). Glycans were placed in the syringe at a concentration of 1–5 mM, and O6 was placed in the cell at a concentration of 54–67.7 μM. The O6 concentration was determined by UV absorbance at 280 nm using calculated extinction coefficients. Experiments were carried out at 25 °C and consisted of 16 injections of 2.45 μL, with injection duration of 1 s, injection interval of 180 s, and reference power of 5 μcal. Fitting of integrated titration peaks was performed with Origin 7.0 software using a single-site binding model to measure the affinity constant (Ka) value. The Kd value was then calculated as the inverse of the Ka.
5
Calorimetric Analysis of Fd-FNR Interaction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Calorimetric experiments were performed basically as described previously 17 except that NaCl was omitted in the reaction (50 mm Tris/HCl, pH 7.5). Protein samples were dialyzed against 50 mm Tris/HCl, pH 7.5, and degassed for 3 min before being loaded into the calorimeter. Calorimetric experiments were performed with an Auto‐iTC 200 instrument (GE Healthcare Biosciences, Chicago, IL, USA) at 298 K. In the injection syringe, 500 μm Fd I was titrated into 50 μm wild‐type or mutant L‐FNR in the ITC cell. Titration experiments consisted of 38 injections spaced at intervals of 150 s (filter period 5 s). The injection volume was 1.0 μL, and the cell was continuously stirred at 1000 r.p.m. Thermodynamic parameters of the complex formation between Fd and FNR were obtained as described previously 18 using the one set of independent binding site model supplied by the microcal origin 7.0 software, OriginLab Corporation, Northampton, MA, USA.
6
Titration of NPM1 Construct
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Titrations were performed using a GE Auto-iTC200 instrument (Malvern, Malvern, UK), at 25 °C, with the NPM1 construct in the cell. Proteins and peptides were dialyzed overnight against the reaction buffer, consisting of 10 mM Na phosphate, 150 mM NaCl, 2 mM DTT, pH 7.0. The concentrations were selected to be below the phase separation threshold and were determined from A280nm of the protein or peptide diluted in 6 M guanidine hydrochloride buffer.
7
Cdk2/Cyclin A Binding Kinetics
8
Copper Binding Affinity of AA13 Lytic Polysaccharide Monooxygenases
9
DAZAP1 Binding Kinetics with hnRNPs
10
Thermodynamics of DJ-1 compound binding
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!